Multiplexed UPLC-MS/MS method for neonatal second-tier screening of inherited metabolic disorders
Salonen, Juho (2023-06-15)
Multiplexed UPLC-MS/MS method for neonatal second-tier screening of inherited metabolic disorders
(15.06.2023)
Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
suljettu
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2023072691366
https://urn.fi/URN:NBN:fi-fe2023072691366
Tiivistelmä
Inborn errors of metabolism (IEM) are a group of disorders that generally involve a failure of metabolic pathways or storage of products of metabolism. These disorders are usually inherited from parents and result from mutations in genes that code the enzymes involved in these metabolism pathways. Since IEMs usually lead to disability or even death if untreated, there are screening programs that detect the disease, and the baby can be treated before symptoms occur. These diseases are often screened with flow injection analysis mass spectrometry (FIA-MS). However, since this method has no separation, false-positive results can occur when the subject has isobaric interferents for the analytes in their blood.1
There is a wide range of different IEMs. In this project, we developed a second-tier screening method for five diseases: maple syrup urine disease (MSUD), Isovaleric Acidemia (IVA), Methylmalonic acidemia (MMA), propionic acidemia (PA), and X-linked adrenoleukodystrophy (X-ALD). Analytes and interferents of these diseases represent compounds such as amino acids, organic acids, carnitines, and lysophosphatidylcholines.1
Waters TQD triple quadrupole mass spectrometer combined with Acquity binary solvent manager UPLC pump system and a high-temperature column heater was used. The final column used for method development was a Phenomenex Kinetex HILIC column with specifications of 2.6μm 100 x 2.1 mm. Two separate methods were finally created both used ACN and water as mobile phase but had different pH for targeting certain groups for better resolution. Separation of analytes was confirmed with standard samples and deuterium-marked standard molecules.
The developed methods successfully separate MSUD, IVA, PA, MMA, and X-ALD primary biomarkers from most of their interferents in a fast, quantitative, and automatable manner. Analytes separated represent a wide range of different molecules; thus, multiplexing is possible, and the introduction of more analytes of other IEMs into this methodology is highly plausible.
There is a wide range of different IEMs. In this project, we developed a second-tier screening method for five diseases: maple syrup urine disease (MSUD), Isovaleric Acidemia (IVA), Methylmalonic acidemia (MMA), propionic acidemia (PA), and X-linked adrenoleukodystrophy (X-ALD). Analytes and interferents of these diseases represent compounds such as amino acids, organic acids, carnitines, and lysophosphatidylcholines.1
Waters TQD triple quadrupole mass spectrometer combined with Acquity binary solvent manager UPLC pump system and a high-temperature column heater was used. The final column used for method development was a Phenomenex Kinetex HILIC column with specifications of 2.6μm 100 x 2.1 mm. Two separate methods were finally created both used ACN and water as mobile phase but had different pH for targeting certain groups for better resolution. Separation of analytes was confirmed with standard samples and deuterium-marked standard molecules.
The developed methods successfully separate MSUD, IVA, PA, MMA, and X-ALD primary biomarkers from most of their interferents in a fast, quantitative, and automatable manner. Analytes separated represent a wide range of different molecules; thus, multiplexing is possible, and the introduction of more analytes of other IEMs into this methodology is highly plausible.